一株伪狂犬病病毒的分离及鉴定

Isolation and identification of a strain of pseudorabies virus

为了从北京市周边郊区某猪场发病仔猪的脑组织中分离到伪狂犬病病毒，试验采用细胞培养、空斑纯化、PCR鉴定、动物感染试验、电镜观察的方法进行分离。结果表明：家兔接种病毒后可引起典型的伪狂犬病症状，继而死亡；PK-15细胞接种病毒出现典型细胞病变，毒价为1×10^8TCID50／mL；电镜观察可见圆形、有囊膜、直径为150～180nm大小的病毒颗粒；PCR反应可扩增出特异性长为I783bp的DNA片段。说明该分离株为猪伪狂犬病病毒野毒株。

To isolate pseudorabies virus from the brain tissue of diseased piglets at a pig farm in the Beijing surrounding suburb, the methods including cell culture, plaque purification, PCR identification, animal infection test, and electron microscope observation were used in the test. The results showed that typical clinical symptoms of pesudorabies were caused in rabbits after infection, followed by death ; typical cytopathogenic effect appeared in porcine kidney cell line after incubation; the titer of the virus was 1 × 10^8 TCID50/mL; round virus particles with envelope and a diameter of 150 nm - 180 nm could be found under electro - microscope, and a specific DNA fragment with 1 783 bp in size could be expanded from the isolated virus DNA using polymerase chain reaction. The results indicate that the isolate was a wild strain of pseudorabies virus